Journal: bioRxiv

Article Title: Loss of STARD7 Impairs Mitochondrial Phospholipid Homeostasis and Contributes to Mitochondrial Myopathy

doi: 10.1101/2025.11.27.691062

Figure Lengend Snippet: Loss of STARD7 disrupts fast-twitch–associated gene expression and fiber-type composition in soleus muscle (A) PCA of RNA-seq data from control (cont) and mKO soleus muscles. Each dot represents an individual animal (n = 3 per group). (B) Volcano plot of differentially expressed genes (DEGs). Differential expression was analyzed using the edgeR package after applying filterByExpr to remove low-expression genes. Genes with |log fold change| > 1 and FDR < 0.05 were defined as DEGs. Fast-twitch–related genes, including Mybpc2 , Actn3 , Pvalb , Mct4 , and Myh4 , were significantly downregulated in mKO. (C) Heatmap of DEGs (FDR < 0.05, |log fold change| > 1) in the soleus. (D) Functional enrichment analysis of DEGs using the clusterProfiler package for Gene Ontology (GO) and WikiPathways. GO terms and pathways with adjusted p < 0.05 were considered significantly enriched. (E) Immunoblot analysis of fast-twitch markers. Each lane represents an independent animal (n = 3). CBB staining confirms equal loading. (F) Representative image of soleus cross-sections stained with isoform-specific myosin heavy chain antibodies. Blue, type I (MyHC-I); red, type IIa (MyHC-IIa); green, type IIb (MyHC-IIb). Fibers lacking these isoforms were classified as type IIx. Scale bar, 200 μm.

Article Snippet: Antibodies used were anti-STARD7 (15191-1-AP), UQCRC2 (14742-1-AP), PVALB (29312-1-AP), MYBPC2 (25257-1-AP), MCT4 (22787-1-AP), MYH4 (20140-1-AP), MYH7 (22280-1-AP), CRLS1 (14845-1-AP), and PGS1 (17149-1-AP) (Proteintech, Rosemont, IL, USA); anti-TAZ (sc-365810) (Santa Cruz Biotechnology, Dallas, TX, USA); OXPHOS complexes (ab110413) (Abcam, Cambridge, UK); anti-OPA1 (612606) (BD Biosciences, San Jose, CA, USA); and 4-HNE (MHN-100P) (JaICA, Shizuoka, Japan).

Techniques: Gene Expression, RNA Sequencing, Control, Muscles, Quantitative Proteomics, Expressing, Functional Assay, Western Blot, Staining

Journal: International Journal of Molecular Sciences

Article Title: G9a Knockdown Suppresses Cancer Aggressiveness by Facilitating Smad Protein Phosphorylation through Increasing BMP5 Expression in Luminal A Type Breast Cancer

doi: 10.3390/ijms23020589

Figure Lengend Snippet: G9a knockdown may lead to increase in BMP5 expression. ( A ) Regulatory factors suppressing tumor aggressiveness were identified by using microarray analysis. ( B ) Expression levels for each gene were compared (ratios between gene expression levels of non-silencing control). In two G9a knockdown cells (shG9a #1 and shG9a #2), expression levels for each gene were compared to those of non-silencing control. Then the most upregulated and downregulated genes were screened. ( C ) G9a knockdown cells showed increase in BMP5 expression. ( D ) Reduced G9a occupancy was found at the BMP5 promoter region. G9a knockdown cells also showed decreased H3K9me2 occupancy at the promoter region of BMP5. p values were calculated using ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Western blot analysis was performed with antibodies specific for the following proteins: G9a (#229455, Abcam, Cambridge, MA, USA), E-cadherin (#3195S, Cell Signaling, Boston, MA, USA), β-catenin (#8480P, Cell Signaling, Boston, MA, USA), ZO-1 (#5406P, Cell signaling, Boston, MA, USA), PARP (#9542S, Cell signaling, Boston, MA, USA), Caspase-7 (#9492S, Cell signaling, Boston, MA, USA), BMP5 (#13253-1-AP, Proteintech, Chicago, IL, USA), p -Smad1/5/9 (#13820T, Cell Signaling, Boston, MA, USA), p -Smad1/5 (#9516T, Cell Signaling, Boston, MA, USA), Smad1 (#6944T, Cell Signaling, Boston, MA, USA) and β-actin.

Techniques: Knockdown, Expressing, Microarray, Gene Expression, Control

Journal: International Journal of Molecular Sciences

Article Title: G9a Knockdown Suppresses Cancer Aggressiveness by Facilitating Smad Protein Phosphorylation through Increasing BMP5 Expression in Luminal A Type Breast Cancer

doi: 10.3390/ijms23020589

Figure Lengend Snippet: BMP5 contributes to reduce migration and invasion abilities of tumor cells. ( A ) Low BMP5 expression is associated with poor survival outcomes of breast cancer patients. ( B ) Patients with stage 3 disease showed higher BMP5 levels than those of stage 2 patients. ( C , D ) Migration/invasion abilities were decreased by treatment of recombinant BMP5; however, the capabilities were enhanced following BMP5 downregulation. p values were calculated using ANOVA. * p < 0.05, ** p < 0.01, **** p < 0.0001.

Article Snippet: Western blot analysis was performed with antibodies specific for the following proteins: G9a (#229455, Abcam, Cambridge, MA, USA), E-cadherin (#3195S, Cell Signaling, Boston, MA, USA), β-catenin (#8480P, Cell Signaling, Boston, MA, USA), ZO-1 (#5406P, Cell signaling, Boston, MA, USA), PARP (#9542S, Cell signaling, Boston, MA, USA), Caspase-7 (#9492S, Cell signaling, Boston, MA, USA), BMP5 (#13253-1-AP, Proteintech, Chicago, IL, USA), p -Smad1/5/9 (#13820T, Cell Signaling, Boston, MA, USA), p -Smad1/5 (#9516T, Cell Signaling, Boston, MA, USA), Smad1 (#6944T, Cell Signaling, Boston, MA, USA) and β-actin.

Techniques: Migration, Expressing, Recombinant

Journal: International Journal of Molecular Sciences

Article Title: G9a Knockdown Suppresses Cancer Aggressiveness by Facilitating Smad Protein Phosphorylation through Increasing BMP5 Expression in Luminal A Type Breast Cancer

doi: 10.3390/ijms23020589

Figure Lengend Snippet: G9a knockdown facilitates Smad protein phosphorylation via BMP5 activation. ( A , B ) G9a-knockdown-induced increase in BMP5 expression had no effect on the total level of either Smad1 or Smad5. G9a knockdown increased phosphorylation of Smad1/5/9. ( C ) ICC demonstrated that nuclear translocation of pSmad1/5/9 was increased in G9a-depleted MCF7 cells. ( D ) Similar tendency was found after treatment of recombinant BMP5. p values were calculated using ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns = not significant.

Article Snippet: Western blot analysis was performed with antibodies specific for the following proteins: G9a (#229455, Abcam, Cambridge, MA, USA), E-cadherin (#3195S, Cell Signaling, Boston, MA, USA), β-catenin (#8480P, Cell Signaling, Boston, MA, USA), ZO-1 (#5406P, Cell signaling, Boston, MA, USA), PARP (#9542S, Cell signaling, Boston, MA, USA), Caspase-7 (#9492S, Cell signaling, Boston, MA, USA), BMP5 (#13253-1-AP, Proteintech, Chicago, IL, USA), p -Smad1/5/9 (#13820T, Cell Signaling, Boston, MA, USA), p -Smad1/5 (#9516T, Cell Signaling, Boston, MA, USA), Smad1 (#6944T, Cell Signaling, Boston, MA, USA) and β-actin.

Techniques: Knockdown, Phospho-proteomics, Activation Assay, Expressing, Translocation Assay, Recombinant

Journal: Obesity (Silver Spring, Md.)

Article Title: Characterization of herpes virus entry mediator as a factor linked to obesity.

doi: 10.1038/oby.2009.250

Figure Lengend Snippet: Figure 1 Data from real-time reverse transcription–PCR showing herpes virus entry mediator (HVEM) gene expression in (a) subcutaneous and visceral human adipose tissue and (b) subcutaneous adipose tissue of lean vs. obese subjects. (c) Western blot analysis of HVEM in subcutaneous human adipose tissue in lean and obese subjects. Relative mRNA levels were normalized to the levels of cyclophilin A and densitometric analyses of HVEM protein were normalized to β-actin. Western blot bands are representative of n = 12 subjects. *P < 0.05.

Article Snippet: Membranes were immunoblotted with antihuman HVEM (Proteintech, Chicago, IL) and β-actin (Santa Cruz Biotechnology, Santa Cruz, CA).

Techniques: Reverse Transcription, Virus, Gene Expression, Western Blot

Journal: Obesity (Silver Spring, Md.)

Article Title: Characterization of herpes virus entry mediator as a factor linked to obesity.

doi: 10.1038/oby.2009.250

Figure Lengend Snippet: Figure 2 Herpes virus entry mediator (HVEM) gene expression in adipocytes. (a) Data from real-time reverse transcription–PCR (RT-PCR) showing HVEM gene expression in stromovascular cells (SVCs) and mature adipocytes (ad) separated from human adipose tissue. Data are means ± s.e.m. (n = 8). (b) Data from real-time RT-PCR and (c) western blot analysis of HVEM in primary culture of human preadipocytes during the differentiation process. Relative mRNA levels were normalized to the levels of cyclophilin A and densitometric analyses of HVEM protein were normalized to β-actin. Data are representative of three different experiments. *P < 0.05.

Article Snippet: Membranes were immunoblotted with antihuman HVEM (Proteintech, Chicago, IL) and β-actin (Santa Cruz Biotechnology, Santa Cruz, CA).

Techniques: Virus, Gene Expression, Reverse Transcription, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR, Western Blot

Journal: Poultry Science

Article Title: In ovo injection of betaine promotes adrenal steroidogenesis in pre-hatched chicken fetuses

doi: 10.1016/j.psj.2022.101871

Figure Lengend Snippet: Nucleotide sequences of specific primers used for real-time PCR.

Article Snippet: Western blot analysis was performed following the protocols provided by the manufacturers, using the primary antibodies for StAR (BS6960, Bioworld, Bloomington, IN, diluted 1:1,200), CYP11A1 (13363-1-AP, Proteintech, Chicago, IL, diluted 1:1,400), HMGCR (BS6625, Bioworld, Bloomington, IN, diluted 1:1,000), SREBP1 (SC-366, Santa Cruz, Dallas, TX, diluted 1:200), GR (Custom made for chickens by Genecreate Biotech Co., Wuhan, China, diluted 1:1,000), CREB (AB31387, Abcam, Cambridge, UK, diluted 1:1,000), BHMT (15965-1-AP, Proteintech, Chicago, IL, diluted 1:1,000), GNMT (18790-1AP, Proteintech, Chicago, IL, diluted 1:1,000), AHCYL1 (10658-3AP, Proteintech, Chicago, IL, diluted 1:1,000), and DNMT1 (24206-1-AP, Proteintech, Chicago, IL, diluted 1:1,000). β-actin (AP0060, Bioworld, Bloomington, IN, diluted 1:10,000 ) and Tublin-β (AP0064, Bioworld , Bloomington, IN, diluted 1:5,000) were used as internal references.

Techniques:

Journal: Cardiovascular diabetology

Article Title: Klotho inhibits renal ox-LDL deposition via IGF-1R/RAC1/OLR1 signaling to ameliorate podocyte injury in diabetic kidney disease.

doi: 10.1186/s12933-023-02025-w

Figure Lengend Snippet: Fig. 6 Klotho regulated podocytic ox-LDL deposition via IGF-1R/RAC1/OLR1 signal axis. A IF analysis of the expression of Nephrin and RAC1 following administration of the inhibitor (PPP) and agonist (IGF2) of IGF-1R on KL+/− DKD and TgKL DKD, respectively. B Western blot analysis was conducted to determine the expression of RAC1, Podocin, and Cleaved Caspase3 following administration of the inhibitor (PPP) and agonist (IGF2) of IGF-1R on KL+/− DKD and TgKL DKD, respectively. C Representative western blot and summarized data showing the effects of administration of the inhibitor (PPP) and agonist (IGF2) of IGF-1R in the absence or presence of Klotho on the relative protein levels of OLR1, RAC1, Podocin, WT1 and Cleaved Caspase3. *P < 0.05; **P < 0.01; ***P < 0.001

Article Snippet: The antibodies used were listed below: anti-ox-LDL antibody (1:100, orb10973, Biorybt), anti-Podocin antibody (1:200, PA5-79757, Invitrogen), anti-Klotho antibody (1:100, PA5-88303, Invitrogen), anti-WT1 antibody (1:50, ab89901, Abcam), anti-PDGFRβ antibody (1:50, ab89901, Abcam), anti-GATA3 antibody (1:2400, ab199428, Abcam), anti-CXCL16 antibody (1:200, 60123- 1-Ig, Proteintech), anti-Parkin antibody (1:100, YT3593, ImmunoWay), anti-RAC1 antibody (1:20, AF385-SP, R&D), anti-Nephrin antibody (1:10, sc-376522, Santa Cruz), anti-OLR1 antibody (1:100, 11837-1-AP, Proteintech), anti-SYNPO antibody (1:200, 21064-AP, Proteintech), anti-IGF-1R antibody (1:10, sc-81464, Santa Cruz), goat polyclonal secondary antibody to mouse Alexa fluor 488 (1:400, ab150113, Abcam), goat polyclonal secondary antibody to rabbit Alexa fluor 555 (1:400, ab150078, Abcam), goat polyclonal secondary antibody to rabbit Alexa fluor 647 (1:400, ab150079, Abcam), goat antimouse Alexa fluor 568 (1:400, ab175473, Abcam), DAPI (1:1000, C1002, Beyotime).

Techniques: Expressing, Western Blot